Chen Yihong, Bi Haitao, Li Xiaoyun, Zhang Zezhi, Yue Haitao, Weng Shaoping, He Jianguo
Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, South China Sea Bio-Resource Exploitation and Protection Collaborative Innovation Center (SCS-REPIC), School of Marine Sciences, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, People's Republic of China.
State Key Laboratory for Biocontrol, OE Key Laboratory of Aquatic Product Safety, Institute of Aquatic Economic Animals and Guangdong Province Key Laboratory for Aquatic Economic Animals, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, People's Republic of China.
R Soc Open Sci. 2017 Apr 26;4(4):160379. doi: 10.1098/rsos.160379. eCollection 2017 Apr.
A previous study found the key transcription factor of PERK-eIF2α pathway cyclic AMP-dependent transcription factor 4 (LvATF4) was involved in the transcriptional regulation of white spot syndrome virus (WSSV) gene . Knocked-down expression of LvATF4 reduced the viral copy number and the cumulative mortality of WSSV-infected shrimp. These results suggested that wsv023 may be critical to WSSV infection but the precise function of wsv023 was still unknown. By using co-immunoprecipitation and pull-down assays, we show that wsv023 interacts with gamma complex-associated protein 2 (LvGCP2), which is the core protein of the γ-tubulin small complex. Knocked-down, the gene significantly reduced the copy number of WSSV in muscle, as well as the cumulative mortality of infected shrimp. And PERK-eIF2α pathway inhibition also showed reduced virus copy number and abrogated shrimp mortality. Furthermore, overexpression of wsv023 inhibited the formation of microtubules in 293T cells. Flow cytometry revealed that WSSV infection similarly decreased the formation of microtubules in haemocytes. These findings suggested that wsv023 plays a role in microtubule organization in host cells, which in turn may be beneficial to WSSV.
先前的一项研究发现，PERK-eIF2α 信号通路的关键转录因子环磷酸腺苷依赖性转录因子 4（LvATF4）参与了对白斑综合征病毒（WSSV）基因的转录调控。敲低 LvATF4 的表达可降低病毒拷贝数以及感染 WSSV 的对虾的累积死亡率。这些结果表明 wsv023 可能对 WSSV 感染至关重要，但 wsv023 的具体功能仍不清楚。通过免疫共沉淀和下拉实验，我们发现 wsv023 与 γ-微管蛋白小复合体的核心蛋白γ复合体相关蛋白 2（LvGCP2）相互作用。敲低该基因可显著降低对虾肌肉中 WSSV 的拷贝数以及感染对虾的累积死亡率。并且抑制 PERK-eIF2α 信号通路也可降低病毒拷贝数并消除对虾的死亡率。此外，wsv023 的过表达抑制了 293T 细胞中微管的形成。流式细胞术显示，WSSV 感染同样降低了血细胞中微管的形成。这些发现表明，wsv023 在宿主细胞的微管组织中发挥作用，这反过来可能对 WSSV 有利。
